Thiophene-linked tetramethylbodipy-labeled nucleotide for viscosity-sensitive oligonucleotide probes of hybridization and protein-DNA interactions
Jazyk angličtina Země Velká Británie, Anglie Médium print-electronic
Typ dokumentu časopisecké články, práce podpořená grantem
PubMed
31919486
DOI
10.1039/c9ob02634g
Knihovny.cz E-zdroje
- MeSH
- DNA vazebné proteiny metabolismus MeSH
- DNA-dependentní DNA-polymerasy metabolismus MeSH
- DNA metabolismus MeSH
- fluorescenční spektrometrie MeSH
- hybridizace nukleových kyselin * MeSH
- kationty MeSH
- lidé MeSH
- lipidy chemie MeSH
- nádorové buněčné linie MeSH
- nukleotidy chemická syntéza chemie MeSH
- oligonukleotidové sondy metabolismus MeSH
- rozpouštědla chemie MeSH
- sekvence nukleotidů MeSH
- sloučeniny boru chemie MeSH
- teplota MeSH
- thiofeny chemie MeSH
- vazba proteinů MeSH
- viskozita MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- 4,4-difluoro-4-bora-3a,4a-diaza-s-indacene MeSH Prohlížeč
- DNA vazebné proteiny MeSH
- DNA-dependentní DNA-polymerasy MeSH
- DNA MeSH
- kationty MeSH
- lipidy MeSH
- nukleotidy MeSH
- oligonukleotidové sondy MeSH
- rozpouštědla MeSH
- sloučeniny boru MeSH
- thiofeny MeSH
Cytosine 2'-deoxyribonucleoside dCTBdp and its triphosphate (dCTBdpTP) bearing tetramethylated thiophene-bodipy fluorophore attached at position 5 were designed and synthesized. The green fluorescent nucleoside dCTBdp showed a perfect dependence of fluorescence lifetime on the viscosity. The modified triphosphate dCTBdpTP was substrate to several DNA polymerases and was used for in vitro enzymatic synthesis of labeled oligonucleotides (ONs) or DNA by primer extension. The labeled single-stranded ONs showed a significant decrease in mean fluorescence lifetime when hybridized to the complementary strand of DNA or RNA and were also sensitive to mismatches. The labeled dsDNA sensed protein binding (p53), which resulted in the increase of its fluorescence lifetime. The triphosphate dCTBdpTP was transported to live cells where its interactions could be detected by FLIM but it did not show incorporation to genomic DNA in cellulo.
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